Process for preparing cardiodilatin fragments; highly purified cardiodilatin fragments and intermediate products for the preparation of same
OwnerHansueli Immer, Wolf-Georg Forssmann +2inventors
InventorsHANSUELI IMMER, WOLF-GEORG FORSSMANN, KNUT ADERMANN, CHRISTIAN KLESSEN
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Abstract
The invention relates to a process for the preparation of cardiodilatin fragments, to highly purified cardiodilatin fragments, and to appropriate intermediates for the preparation of said fragments. Furthermore, the invention relates to highly purified cardiodilatin fragments which are free of peptide impurities and exhibit a single migration peak in capillary electrophoresis, as well as to appropriate processes for the preparation of same.
Claims20
A process for preparation of cardiodilatin fragments of formula I
- R1-ANP(105-121)-R2 (I),
- having a chain length of 17-37 amino acid in total, wherin ANP(105-121) represents the amino acid sequence [SEQ ID NO. 1],
- R1 represents an amino acid chin of sequence ANP(90-104) [SEQ ID NO. 2] or fragments thereof having a chain length of 0-15 amino acids, and
- R2 represents an amino acid chain of sequence ANP(122-126) [SEQ ID. 3] or fragments thereof having a chain length or 0-5 amino acids, characterized in that synthesis is effected via condensation of at lease three partial fragments, the condensation of said partial fragments to give the cardiodilatin fragment of formula I being carried out between the amino acid positions Gly 108 and Arg109 and amino acid positions Gly120 and Cys121.
A process for the preparation of high-purity cardiodilatin fragments R1-ANP(105-121)-R2 having a chain length of 17-37 amino acids in total, wherein R1 represents an amino acid chain of sequence ANP(90-104) or fragments thereof having a chain length of 0-15 amino acids, and R2 represents an amino acid chain of sequence ANP(122-126) or fragments thereof having a chain length of 0-5 amino acids, characterized in that purification of the crude product is performed using a reversed-phase HPLC column, and the cardiodilatin fragment is eluted with a buffer system containing triethylammonium phosphate and acetonitrile.
High-purity cardiodilatin fragments R1-ANP(105-121)-R2 having a chain length of 17-37 amino acids in total, wherein R1 represents an amino acid chain of sequence ANP(90-104) or fragments thereof having a chain length of 0-15 amino acids, and R2 represents an amino acid chain of sequence ANP(122-126) or fragments thereof having a chain length of 0-5 amino acids, characterized in that they are substantially free of peptide impurities and exhibit a single migration peak in the purity analysis using capillary electrophoresis.
Peptide fragments having the amino acid sequence R1-ANP(105-108), wherein R1 represents an amino acid chain of sequence ANP(90-104) or fragments thereof having a chain length of 0-15 amino acids, as well as their derivatives modified by protecting groups.
Peptide fragment having the amino acid sequence ANP(109-120), as well as derivatives thereof modified by protecting groups.
Peptide fragments having the amino acid sequence ANP(109-121)-R2, wherein R2 represents an amino acid chain of sequence ANP(122-126) or fragments thereof having a chain length of 0-5 amino acids, as well as their derivatives modified by protecting groups.
Peptide fragments having the amino acid sequence Cys121-R2, wherein R2 represents an amino acid chain of sequence ANP(122-126) or fragments thereof having a chain length of 3-5 amino acids, as well as their derivatives modified by protecting groups.
Dependencies are inferred from the claim text.